CELL SIGNALING

Frederic A. Montaño, MD, PTRP, DPBO

 Cell-to-Cell Communication

• Ligand binds to
receptors in the plasma
membrane, cytoplasm
or nucleus

• Receptors interact with

1 or more intracellular
messengers going into
activation/deactivation

• Signaling proteins
interact with target
proteins which regulate
cell growth and
division
 Cell-to-Cell Communication
Characteristics

• Multiple, hierarchical
steps

• Amplification of the
hormone-receptor binding
event

• Activation of multiple
pathways and regulation of
multiple cellular functions

• Antagonism by constitutive
and regulated feedback
mechanism
 Cell-to-Cell Communication
• Secretion of signaling
molecules is cell type
specific

• Ability to respond to
a specific signaling
molecule depends on
the expression of
receptors that bind
the signaling molecule
with high affinity and
specificity
 Cell-to-Cell Communication

• Signaling molecules
can act over long or
short distances and
require cell-to-cell
contact or very close
proximity
 Contact Dependent Signaling

• Important during
development and in
immune response
 Paracrine Signaling

• Released by one type

of cell and act on
another type

• Usually taken up by
target cells or rapidly
degraded
 Autocrine Signaling

• Release of a molecule
that affects the same
cell or other cells of
the same type
 Synaptic Signaling

• Neurons transmit electric

signals along the axons
with release of
neurotransmitters at
synapses that affect the
function of other neurons
or cells distant from the
neuron cell body
 Endocrine Signaling

• Hormones are
secreted into the
blood supply and
widely dispersed into
the body

• Relatively slow (sec.

to mins.)
 Gap Junction Signaling
• Forms between adjacent
cells

• Allows small intracellular

signaling molecules (<1200
Da) to diffuse from
cytoplasm of one to an
adjacent cell

• Permeability is regulated by
cytosolic Ca++, H+, cAMP
& membrane potential

• Seen in cardiac and smooth

ms. cells
 How Fast is the Response

• Depends on the
mechanism of
delivery

• Depends on the ability

of the molecule to
reach a particular cell,
on expression of the
cognate receptor & on
cytoplasmic signaling
molecules that interact
with receptor
 Receptors

2 Basic Classes

1. Plasma Membrane
receptors

2. Nuclear receptors
 Plasma
Membrane
Receptors
1. Ion channel linked receptors
2. G protein coupled receptors
(GPCRs)
3. Catalytic receptors
4. "Pseudo receptor" that undergo
regulated intramembrane
proteolysis (RIP)
 Ion Channel-linked Receptors

• A.k.a ligand gated

ion channels

• Mediate direct & rapid

synaptic signaling
between electrically
excitable cells
 GPCRs

• Regulate the activity

of other proteins
like enzymes and
ion channels

• Interaction is
mediated by
heterotrimeric G
proteins composed
of alpha, beta &
gamma subunits
 Catalytic Receptors

• A.k.a enzyme
linked receptors

• Ex. protein kinases

which can
phosporylate a
specific subset of
proteins or amino
acids which in turn
activates or inhibit
protein activity
 RIPs
• Do not fit the classic
definition of receptors

• On activation by the ligand,

membrane proteins undergo
RIProteolysis which
elaborates a cystolic peptide
fragment that enters the
nucleus & regulate gene
expression

• Ex. sterol element binding

protein (SREB)
 Nuclear Receptors

• Where small hydrophobic

molecules bind (steroid hormones,
thyroid hormones, retinoids, Vit.
D, cortisol, aldosterone)

• Receptors can be found in the

cytosol or bound to the DNA in
the nucleus but inactive

• Activated once the binding of a

hormone dissociates the
inhibitory complex

• Hormone receptor complex binds

to DNA and regulates gene
transcription
 Signal Transduction Pathways

• Plasma membrane
receptors: intracellular
signaling pathways (2nd
messengers)

• Nuclear receptors:
regulation of gene
expression
 2nd Messengers

• Small molecules: cAMP, cGMP, Ca

++, DAG

• Relay the signal from one protein to

another/from one region of the cell
to another

• Produce large amounts of additional

signaling molecules

• Convert the signal into different form

• Act as molecular switches

• Can adjust the signal sensitivity by

adaptation/ desensitization
 Ion Channel Linked Signal
Transduction Pathways

• Transduces a chemical
signal into an electrical
signal

• Ex. RyR receptor

 G Protein Coupled Signal
Transduction Pathways

• G proteins

• heteromeric proteins
composed of 3 subunits

• α, β and γ

• can assemble into

hundreds of different
combinations
 G Protein Coupled Signal
Transduction Pathways

• If ligand is absent, G
proteins are inactive
where GDP binds to α
subunit

• Receptor is activated
once ligand is bound and
GTP binds to α subunit
 G Protein Coupled Signal
Transduction Pathways
• GTP binding-> dissociation
of α subunit from the complex
& results in release of α
subunit from βγ dimer

• G proteins:

• activated by guanine
nucleotide exchange factors
(GEFs)

• inactivated by GTPase-
accelerating proteins (GAPS)
 Termination of GPCR Signal
Transduction Pathways
• Hydrolysis of GTP to GDP and Pi

• Facilitated by RGS proteins which

inactivates the signal

• Desentization & endocytic

removal of receptors from the
plasma membrane

• Mediated by GPCR kinases

(GRKs)/β arrestin which down
regulates a response during
prolonged exposure to elevated
hormone levels
 G Proteins Acting via Adenylyl
Cyclase
• Receptor with ligand interacts
with G protein bound to α
subunit of the αs class ->
adenylyl cyclase activation ->
inc. cAMP levels -
>activation of protein
kinase A (PKA)

• PKA can enter cell and activate

cAMP response element-
binding (CREB) protein ->
gene transcription

• cAMP degraded to AMP by

cAMP phosphodiesterases
 G Proteins Acting via
Phosphodiesterase
• Absorption of light by
rhodopsin in the eye's rods -
>activation of G protein
transducin -> activation
of cGMP
phosphodiesterase->
lowers conc. of cGMP ->
closure of cGMP activated
cat-ion channel-> alteration
of membrane voltage

• Reason why we can see

with our eyes: sensitivity
of rhodopsin to light
stimulus
 G Proteins Acting via Phospholipase

• G protein coupled to αq
subunit stimulates
phospholipase C -> converts
phosphatidyleinositol 4,5
biphosphate (PIP2) to 1,4,5-
inositol triphosphate (InsP3)
and DAG

• InsP3: 2nd messenger;

releases Ca++ into cytosol

• DAG: activates protein

kinase C (PKC)
 Arachidonic Acid Signaling Pathway

• Phospholipase A2: releases

arachidonic acid

• Arachidonic acid: stimulates

inflammation

• 3 pathways involved:

• Cyclooxygenase pathway

• Lipooxygenase pathway

• Epoxygenase pathway
1st Pathway: Cyclooxygenase (COX)
Pathway
• Cyclooxygenase: facilitate the
metabolism of arachidonic acid to
prostaglandin, thromboxanes &
prostacylins

• Prostaglandin: platelet
aggregation, airway constriction
& inflammation

• Thromboxanes: platelet
aggregation, vasoconstriction

• Prostacylin: inhibit platelet

aggregation, vasodilatation
 2nd Pathway: 5- Lipoxygenase
Pathway

• 5-Lipoxygenase: conversion of
arachidonic acid to leukotrienes

• Leukotrienes: allergic anc &

inflammatory responses like
asthma, RA, inflammatory
bowel disease
3rd Pathway: Epoxygenase Pathway

• Epoxygenase: facilitaes
generation of
hydroxyeicosatetrenoic acid
(HETE) and cis-
epoxyeicosatrienoic acid
(EET)

• HETE & EET: increase Ca++

release from ER & stimulate
cell proliferation
 Protein Phosphatases & Phosphodiesterases
Reverse the Action of Cyclic Nucleotide Kinases

• Signal termination started

by cAMP & cGMP:

• Phosphodiesterases:
breakdown of cAMP &
cGMP to AMP & GMP

• Phosphatases:
dephosphorylate effector
proteins which were
phosphorylated by
kinases such as PKA
 Small Monomeric G Proteins
• Single 20-40 kDa protein;; membrane bound

• Activity depends on binding of GTP; regulated by GEFs & GAPs

• Ras GTPase: regulate gene expression, cell proliferation,

differentation, and survival

• Rho GTPase: regulate actin cytoskeletal organization, cell cycle

progression & gene expression

• Rab GTPase: regulate intravascular transport & trafficking of

proteins

• Ran GTPase: regulate nucleocytoplasmic transport of RNA &

proteins

• Arf GTPase: regulate vesicular transport

 Catalytic Receptor Linked Signal
Transduction Pathways

• 4 Classes of Receptors:

1. Receptor guanylyl cylases

2. Receptor serine/threonine kinases

3. Receptor tyrosine kinases (RTks)

4. Tyrosine-kinase associated receptors

 Receptor Guanylyl Cyclases

• ANP binds to receptor --> receptor

dimerization--> activation of
guanylyl cyclase-->metabolize
GTP to cGMP--> activates cGMP
dependent protein kinase (PKG)

• Ex: ANP inhibits sodium & water

reabsorption by the collecting duct

• NO activates guanylyl cyclase

receptor which relaxes smooth
ms in coronary arteries

• Seen also as the mechanism if

nitroglycerin
Receptor Serine/Threonine Kinases

• TGF-Β receptor: 2
subunits

• Binding of TGF-B to type

II subunit induces it to
phosphorylate the type I
subunit-->elicits cellular
response
 Receptor Tyrosine Kinase

• Nerve growth factor (NGF)

• Ligand binding to 2 NGF receptor--

> dimerization & activation of
tyrosine kinase activity

• Insulin receptor

• Tetrameric: 2α & and 2β subunits

• Binding of insulin to α subunits--

>interaction between 2 α & Β
subunits-->autophosporylation of
tyrosine residues in the catalytic
domains of the Β subunits--
>initiate cellular effects
 Tyrosine Kinase Associated
Receptors

• No intrinsic kinase
activity but associated
with proteins with tyrosine
kinase activity (Src family
& Jannus family (JAK)

• Assembles into
homodimers, heterodimers
or heterotrimers
 Nuclear Receptor-Linked Signal
Transduction Pathways
• Subfamilies:

1. Steroid receptors

2. Receptors that bind

retenoic acid, thyroid
hormones, & Vit D

• Activates transcription factors-->

bind to DNA--> regulate gene
expression

• Can also act as transcriptional

repressors as seen in
glucocorticoids
 Nuclear Receptor-Linked Signal
Transduction Pathways
• Glucocorticoid &
mineralocorticoid receptors

• Located in cytoplasm

• Interact with chaperones (heat

shock proteins)-->
conformational change-->
dissociation of chaperone from
receptor --> facilitate
translocation of hormone bound
receptor to nucleus

• Glucocorticoids suppress
transcription activator protein-1
(AP-1) & nuclear factor kB (NF-
KB) which reduce inflammation
 Nuclear Receptor-Linked Signal
Transduction Pathways

• Estrogen & progesterone

receptors

• Located in nucleus

• Thyroid hormones &

retinoic acid receptors

• Nucleus bound to DNA